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Development of a sodium dodecyl sulfate‐polyacrylamide gel electrophoresis reference method for the analysis and identification of fish species in raw and heat‐processed samples: A collaborative study
Author(s) -
Piñeiro Carmen,
BarrosVelázquez Jorge,
PérezMartín Ricardo I.,
Martínez Iciar,
Jacobsen Tonna,
Rehbein Hartmut,
Kündiger Rainer,
Mendes Rogerio,
Etienne Monique,
Jerome Marc,
Craig Anne,
Mackie Ian M.,
Jessen Fleming
Publication year - 1999
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(19990601)20:7<1425::aid-elps1425>3.0.co;2-r
Subject(s) - chromatography , sodium dodecyl sulfate , chemistry , ultrafiltration (renal) , polyacrylamide gel electrophoresis , extraction (chemistry) , capillary electrophoresis , urea , rnase p , gel electrophoresis , biochemistry , enzyme , rna , gene
A collaborative study was carried out in seven European labs with the aim of achieving a sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) standard operation procedure to identify fish species in raw and cooked samples. Urea and SDS‐containing solutions were evaluated as extractants. Several preelectrophoretic operations — such as treatment with RNase/DNase, ultrafiltration and desalting — and up to ten types of gels and three SDS‐PAGE systems were considered. The SDS‐containing solution allowed a higher protein extractability than urea. Unlike urea extraction, SDS extraction seemed not to be influenced so much by the state of the sample (raw, cooked at 60 o C, cooked at 85 o C). Desalting, ultrafiltration or treatment with RNase/DNase did not improve the discriminatory power of the protein patterns. Commercial homogeneous 15% ExcelGels, especially when they were silver stained, yielded good results and afforded higher reproducibility, thus allowing a better matching of results among the laboratories participating in this collaborative study. Under the optimized technical conditions described above, all the fish species tested, either raw and cooked, yielded reproducible and discriminant species‐specific protein patterns.